Mechanism of NK cell activation induced by coculture with dendritic cells derived from peripheral blood monocytes.

Dendritic cells (DCs) have been regarded as one of the effective antigen-presenting cells, but the relationship between DCs and lymphocytes, in particular natural killer (NK) cells, remains unclear. In this study, we evaluated how DCs interact with both lymphocytes and NK cells using a coculture system. The number of lymphocytes increased significantly when cocultured with DCs (1.8-fold increase). In particular, the proliferation of NK cells was prominent. Furthermore, the coculture of DCs with lymphocytes induced a marked increase in IL-12 and IFN-gamma secretion. When contact between the DCs and lymphocytes was prevented, the secretion of both IL-12 and IFN-gamma was markedly reduced. IFN-gamma production was completely blocked by an anti-IL-12 antibody, indicating that IFN-gamma secretion was dependent on IL-12 secretion. The stimulating effect of the DCs on the proliferation of the lymphocytes was partially suppressed by anti-IL-12 antibodies, and was completely attenuated when cellular contact was prevented. Furthermore, the NK cell proliferation induced by coculture with DCs was significantly blocked by the inhibition of the interaction of either CD40-CD40L or CD28-B7 molecule. The coculture with DCs enhanced NK activity by 40%, and this was partially suppressed by anti-IL-12 antibodies and was completely blocked by the inhibition of cell-to-cell contact. These results indicate that the activation of NK cells by DCs is partially mediated by IL-12 secretion, and that direct contact between DCs and NK cells play a major role in this response.

Prevention of murine acute graft-versus-host disease by staphylococcal enterotoxin B treatment.

Retroviral superantigens such as minor lymphocyte stimulating (Mls) antigen play an important role in the pathogenesis of acute graft-versus-host disease (GVHD). However, it remains unclear how exogenous bacterial superantigens modulate acute GVHD. In this study, we tested the effects of staphylococcal enterotoxin B (SEB) on the development of acute GVHD in a model involving the systemic transfer of parental C57Bl/6 spleen cells into BDF1 mice. SEB treatment suppressed the expansion of donor-derived T cells and blocked the decrease in the number of host cells. Impaired haematopoiesis was actually rescued by treatment with SEB. In SEB-treated mice, both spontaneous proliferation and IL-2 production in T cells were suppressed on day 2 after parental cell infusion. On day 21, the number of donor-derived CD4+ Vbeta8+ T cells markedly decreased in the spleen of SEB-treated mice. Donor-derived CD4+ T cells failed to proliferate in response to host alloantigens, and both donor- and host-derived T cells were unable to produce IL-2 in response to concanavalin A stimulation, suggesting that SEB treatment induced a general immunosuppressive state. Our results indicate that SEB treatment prevents the development of acute GVHD by leading to unresponsiveness of donor-derived T cells against host alloantigens in a Vbeta-restricted and unrestricted manner.

Low molecular weight dextran attenuates increase in extravascular lung water caused by ARDS.

We studied the effect of low molecular weight dextran (mean molecular weight 40,000, Dextran 40; LMD) on the accumulation of extravascular lung water (EVLW), and also on hemodynamics and blood gases, in the oleic acid (OA)-injured lung in pentobarbital anesthetized rats. Starting just before the OA injection (0.01 mL/kg via femoral vein), 10% LMD in lactated Ringer's solution was infused throughout the experiment (5 mL/kg/h) instead of lactated Ringer's solution. OA caused acute lung injury leading to decreased oxygenation (PaO2: 87 +/- 11 mmHg versus control group 128 +/- 11) and an increased permeability of the alveolar-capillary membrane, as shown by increases in EVLW (4.89 +/- 0.54 versus control group 4.07 +/- 0.14), and albumin leakage (0.043 +/- 0.015 versus control group 0.010 +/- 0.004). LMD protected against the increase in EVLW (4.14 +/- 0.10) and the hypoxemia (112 +/- 19 mmHg), but it did not reduce the albumin leakage into the alveolar space (0.052 +/- 0.009). These data suggest that LMD may limit the fluid accumulation that is secondary to OA-induced lung injury.

[Significance of product documents (tenpubunsho) cited in judicial precedents in the field of anesthesiology].

The significance of the product documents (Tempubunsho) was mentioned in 21 out of 94 medical malpractice cases published in several laws reports 1963-97. The number of such cases has been increasing in recent years. Among the items described in the product documents, the precautions for use were most frequently mentioned (12 cases) followed by the instructions for administration and dosage (3 cases). In most cases, the judgment was against the medical institution involved due to the violation of legal obligations with respect to the contents of the product documents. Product documents will be taken much more seriously in future lawsuits. Attention should be focused on the contents of the product documents, especially when revised. When the administration of a drug deviates from the contents of the product document, medical evidence is required to support this deviation.

Improvement of an impaired chemiluminescence response to formyl-methionyl-leucyl-phenylalanine in neutrophils from patients with non insulin dependent diabetes mellitus by recombinant human granulocyte-colony stimulating factor.

To explore the possibility that b type recombinant human granulocyte-colony stimulating factor (rhG-CSF) is a useful drug to prevent the morbidity and mortality caused by infections in diabetic patients, we have studied effects of rhG-CSF on chemiluminescence amplified by a luciferin analog (CLA-DCL) and luminol (L-DCL) in response to formyl-Methionyl-Leucyl-Phenylalanine (fMLP) in neutrophils from patients with non insulin dependent diabetes mellitus (NIDDM) (diabetic neutrophils) and healthy subjects (control neutrophils). Both CLA-DCL and L-DCL in diabetic neutrophils were significantly reduced, and L-DCL was more sensitive to this suppression than CLA-DCL. RhG-CSF did not change the basal chemiluminescence in control and diabetic neutrophils, but it primed CLA-DCL and L-DCL. Although, in diabetic neutrophils, the priming effect of rhG-GSF on both CLA-DCL and L-DCL was less compared to that in control neutrophils, L-DCL was more sensitive to this priming effect than CLA-DCL. Because bacterial infection is still an important cause of the morbidity and mortality in diabetic patients, these data suggest that rhG-CSF is a useful drug to prevent the aggravation of bacterial infection in patients with NIDDM.

Sodium butyrate blocks interferon-gamma (IFN-gamma)-induced biosynthesis of MHC class III gene products (complement C4 and factor B) in human fetal intestinal epithelial cells.

Human intestinal epithelial cells have been established as local sites for complement biosynthesis. In this study, we investigated the effects of IFN-gamma and sodium butyrate on biosynthesis of MHC class III gene products (complement C4 and factor B) in the human fetal intestinal epithelial cell line INT-407. IFN-gamma induced a dose- and time-dependent increase in C4 and factor B secretion. However, sodium butyrate dose-dependently inhibited IFN-gamma-induced C4 and factor B secretion. These effects were also observed at the mRNA level. Immunoblotting indicated that IFN-gamma induced a rapid activation of Stat1alpha, and fluorescence immunohistochemistry detected a translocation of Stat1alpha into the nucleus within 1 h. However, the translocation of Stat1alpha was not affected by the addition of sodium butyrate. Nuclear run-on assay indicated that IFN-gamma induced a weak increase in the transcription rate of factor B gene, and sodium butyrate did not affect this response. IFN-gamma and sodium butyrate induced a counter-regulatory effect on C4 and factor B secretion: IFN-gamma acted as a potent inducer, but sodium butyrate potently abrogated these responses. These are mainly regulated through the post-transcriptional mechanism.

[Judicial judgements on anesthesia malpractice in Japan].

We reviewed 75 judicial precedents on anesthetic malpractice during surgical procedure which had appeared in legal journals in the period between 1963 and 1997. Anesthetic techniques employed were: general anesthesia (35 cases), spinal anesthesia (19 cases), local anesthesia (12 cases), and others (9 cases). Anesthesiologists were involved in 16 lawsuits, of which anesthesiologists lost 6 suits between 1986 and 1995. There were 8 cases classified as to be caused by respiratory problems including 2 cases of wrong gas supply. The defendants lost all the 8 cases. On the other hand, the plaintiff lost all the cases of malignant hyperthermia (n = 7). There is a tendency of increase in law suit with general anesthesia. Recent judgments suggested the importance of anesthetic managements, correct recording and appropriate monitoring by anesthesiologist during and immediately after surgery. Spinal anesthesia should be performed by anesthesiologist, and the frequency of anesthetic accident should be decreased. Japan is still in short of anesthesiologists and efforts should be paid to increase the number of anesthesia specialists.

[Effective pentostatin-based treatment of adult T cell leukemia in a patient with severe arthritis].

We report a 48-year-old woman with adult T-cell leukemia who had refractory arthralgia, intense headaches, and fever. Leukemic cell infiltration of the cerebrospinal fluid was detected but no other acute signs were observed. Abnormal lymphocytes with lobulated nuclei were found in the synovial fluid, and a histologic examination revealed proliferation into the synovium. Because combination chemotherapy did not elicit a favorable response, the patient was treated with a pentostatin bolus injection. The articular symptoms disappeared and complete remission was obtained. Six months later, she experienced arthralgia again together with a gradual increase of abnormal lymphocytes in peripheral blood. Sixteen months later, the patient was given pentostatin and achieved a complete remission again. She is still free from relapse without further therapy after 36 months, and her articular symptoms have not returned either. There were no adverse effects due to pentostatin. The patient's serum IL-6 level was elevated, suggesting that IL-6 may play a role in arthropathy.

[Perioperative management for nephrectomy in a long-term hemodialysis patient with anticoagulants for coronary stent].

A 53-year old man on long-term hemodialysis (HD) with anticoagulant therapy was scheduled for nephrectomy due to renal cell carcinoma. Two months before surgery, a coronary stent had been placed due to right coronary artery disease. One week before surgery, percutaneous transmural coronary angioplasty (PTCA) was performed for unstable angina. Aggressive oral antiplatelet therapy (aspirin and ticlopidine) was absolutely required to maintain patency. Following withdrawal of the antiplatelets, unfractionated heparin (UFH) was titrated to an activated partial thromboplastin time (APTT) of 1.5 times greater than the control value. Maintenance UFH (800 U.h-1) was continued until the time of arrival in the operation room (activated clotting time (ACT) was 166 seconds). One hour after arrival, reduced dose of UFH (200 U.h-1) was reinfused, and ACT was 121 140 seconds. Hemodynamic change was minimized using balanced general anesthesia (nitrous oxide-isoflurane, fentanyl, midazolam and vecuronium) accompanied by nitroglycerin and diltiazem. There was no ischemic change on ECG or transesophageal echocardiography. Following surgery, the UFH dose was augmented (400 U.h-1), and the maintenance dose was attained 11 hours after surgery. HD on the second postoperative day was performed uneventfully. This hemodynamic stability might be come from the no water removal. Fourteen days after surgery, the patient was discharged without hemorrhagic complications or clinical ischemic events. We conclude that perioperative UFH infusion is not contraindicated for dialysis patient if strict ACT control is maintained.

Thyroxine-induced production of superoxide anion by human alveolar neutrophils and macrophages: a possible mechanism for the exacerbation of bronchial asthma with the development of hyperthyroidism.

Bronchial asthma worsens after the development of hyperthyroidism. However, the biochemical mechanism of this phenomenon, which is induced by thyroxine (T4), remains obscure. In the present study, we showed that T4 directly stimulates production of superoxide anion by alveolar neutrophils and macrophages. These cells, when sensitized with a patient's serum and then treated with both the patient's specific allergen and T4, produced higher amounts of superoxide anion than did sensitized cells treated with the specific allergen or T4 alone. Our data suggest that T4 enhances production of reactive oxygen species by alveolar neutrophils and macrophages, which might play an important role in the exacerbation of bronchial asthma with the development of hyperthyroidism.

[A reused suction catheter was broken and produced a tracheobronchial foreign body].

We present a rare case involving a broken suction catheter that became lodged in the tracheobronchial tree. An eight-month-old infant was scheduled for hernioplasty. Following intubation with a 4 French (Fr.) endotracheal tube, a 5 Fr. reused suction catheter was applied for suctioning a moderate amount of secretion. This catheter had been resterilized by ethylene oxide gas (EOG). Insertion of the catheter into the tube was not done smoothly, and we could not obtain any secretion. During the extraction of the suction catheter by force, the catheter broke. The distal fragment (20 cm length) seemed to have been lodged in the tracheobronchial tree. Prior to its removal by bronchoscopy, the endotracheal tube was extubated. Fortunately the remaining part of the catheter come out with the tube. Ten cm of the fractured catheter was included in the tube and 5 cm protruded from the tube. The catheter of smaller diameter is easy to be broken even by weaker force. After resterilization by EOG for once, there was no change in length and force at breaking point. Elongation of the broken catheter (85.5%) was less compared with the new sample (155%). At the breaking point, half of the cross section was very smooth and looked as if it had been cut by a razor, while the other half appeared to have been broken by pulling. The break may have started from the crack which had occurred at the insertion or resterilization. Therefore, we should restrict the reuse of small suction catheters, and should always utilize the catheter of the largest size possible.

Effects of antiallergic drugs on the proliferation of estrogen-sensitive mouse Leydig cell line.

The proliferation of B-1F cells established from an estrogen-sensitive mouse Leydig cell tumor is negatively regulated by leukotrienes, whose production is suppressed by the addition of estrogen. Disodium cromoglycate, tranilast, repirinast, tazanolast and pemirolast potassium are used as therapeutic agents for allergic diseases. They are known to inhibit the release of chemical mediators such as histamine and leukotrienes from peripheral leukocytes. It is therefore of interest to determine whether these drugs affect the proliferation of B-1 F cells. The drugs in the culture medium stimulated the proliferation of B-1F cells to various extents. The results suggest that chronic administration of these drugs to the patients with allergic diseases may possibly enhance the proliferation of some kinds of tumor cells, resulting in the change from latent to clinical tumors, although there are significant differences between in vitro cultured animal cells and in vivo humans.

Effects of osmotic pressure on intrathecal and epidural lidocaine anesthesia.

Lidocaine (1%), either in plain distilled water or in 10% dextrose, was intrathecally or epidurally administered to urethane-chloralose anesthetized cats. Electrical stimulation was applied to the gracile tract at a cervical level, and the resultant antidromic compound action potentials were recorded from the sural nerve. Lidocaine dissolved in plain distilled water was more effective than lidocaine dissolved in 10% dextrose solution in suppressing the compound action potentials. Lidocaine-free plain distilled water or dextrose solution caused partial suppression of the compound action potentils. The suppression was more marked following plain distilled water application than following application of 5% or 10% dextrose.

[Minimizing of the blood preparation for elective surgery by maximum surgical blood order schedule as well as type and screen--two year's experience in Nagahama City Hospital].

To make the most of the blood products, we have introduced maximum surgical blood order schedule (MSBOS) as well as Type and Screen (T&S) into Nagahama City Hospital. As the hospital dose not have blood transfusion service unit, we, anesthesiologists set up MSBOS and the procedure of blood preparations for elective surgery. The results of two year's experience demonstrate that there was no trouble by reducing the blood preparation. The cross-match to transfusion ratio (C/T ratio) in surgical procedures and that of red cell products in our hospital have decreased, the effective use of red cell products has been promoted, and the outdating of the blood product has been reduced.

Comparison of responses of canine pulmonary artery and vein to angiotensin II, bradykinin and vasopressin.

Responses to angiotensin II, bradykinin and arginine vasopressin were compared in helical strips of canine pulmonary arteries and veins. Angiotensin II contracted the artery but relaxed the vein strip. The artery contraction was augmented by indomethacin and aspirin and was abolished by losartan. The vein relaxation was not affected by endothelium denudation but was abolished by the cyclooxygenase inhibitors, a prostaglandin I2 synthase inhibitor and losartan. The bradykinin-induced artery relaxation was inhibited by endothelium denudation, NG-nitro-L-arginine (L-NA) or indomethacin and abolished by their combined treatment. The vein relaxation produced by bradykinin was endothelium-independent and was abolished by indomethacin. Vasopressin produced a slight relaxation in the arteries, which was abolished by endothelium denudation and L-NA. The vein relaxation produced by vasopressin was abolished by endothelium denudation and combined treatment with L-NA and indomethacin. It may be concluded that (1) activation of angiotensin AT1 receptor subtype in smooth muscle produces contraction and also relaxation due to prostaglandin I2 release; the former predominates over the latter in the artery, whereas only the latter is operative in the vein, (2) the bradykinin-induced relaxation is due to nitric oxide (NO) from the endothelium and prostaglandin I2 from subendothelial tissues in the artery and solely to prostaglandin I2 in the veins, and (3) the vasopressin-induced relaxation is mediated by endothelial NO in the artery, and NO and prostaglandin I2 in the vein.

[Perioperative management of dialysed patients for orthopedic surgery].

Recently, renal osteodystrophy is a remarkable problem in patients on long-term hemodialysis (HD). In this retrospective study, we evaluated the perioperative management of 21 patients receiving orthopedic surgery between January 1990 and December 1992. These patients had been maintained on HD for an average of 8.6 years (range, 18 months-20 years). The primary causes of orthopedic surgery were amyloidosis, diabetic gangrene, rheumatoid arthritis and fractures. Laminectomy, replacement of arthropathy, osteosynthesis and amputation of the lower extremity were undertaken. General anesthesia was performed on six patients. Vecuronium was given to all of these patients. Isoflurane was used in 5 patients and sevoflurane in 1 patient. Regional anesthesia was used in 15 patients. During anesthesia, the average infusion rate of intravenous fluids was 2.7 ml.kg-1.h-1, and the intraoperative complications included hypertension in 16, hypotension in 12, arrhythmia in 4 and prolonged sedation in 2 patients. Postoperative complications included hyperkalemia in 2, pneumonia in 2, psychological disorder in 3, clotting fistula in 1 and delayed wound healing in 7 patients. One early death in a diabetic patient following amputation occurred on the 13th postoperative day. Preoperative HD was performed within 24 hours and postoperative HD within 72 hours of the operation. Nafamostat mesilate was used as an anticoagulant. Excessive removal of potassium must be avoided during preoperative HD to prevent arrhythmia. The well-managed elective patients gave a good result. However, extreme care in nutrition and infection control should be taken, especially in diabetic patients.

Comparative study of the mammalian liver innervation: an immunohistochemical study of protein gene product 9.5, dopamine beta-hydroxylase and tyrosine hydroxylase.

The liver innervation of eight different mammalian species was examined by immunohistochemical localization of protein gene product (PGP) 9.5 to visualize the general innervation for autonomic nerve fibres. In addition, dopamine beta-hydroxylase (DBH) and tyrosine hydroxylase (TH), two enzymes involved in catecholamine synthesis, were localized immunohistochemically to delineate hepatic sympathetic nerve fibres. We found that: (1) Within the interlobular region of each species, PGP 9.5, DBH and TH-positive nerve fibres were all seen in close association with branches of hepatic arteries, portal veins and bile ducts. (2) Within the parenchyma of the guinea-pig, cat, dog, pig, monkey and human liver, the presence of the three immuno-positive nerve fibres could be unequivocally identified, although the density of these intralobular fibres showed marked species variation. Moreover, immunoelectron microscopic study confirmed that PGP 9.5-positive nerve terminals of the human liver are in close apposition to hepatocytes. (3) In mouse and rat, no parenchymal nerve fibres immunoreactive for PGP 9.5, TH or DBH could be demonstrated.

[The influences of local anesthetics on ryanodine-induced contracture in rat skeletal muscle].

One hundred and fifty paired extensor long digital muscles were excised from Wistar rats and each muscle was prepared in Krebs-Ringer's solution (K-R solution) then gassed with a mixture of 95% O2-5% CO2. The medium for the control muscles was replaced with K-R solution containing 10(-6) M ryanodine and that for the experimental muscles was replaced with medium containing 10(-6) M ryanodine and local anesthetic (LA) (procaine, tetracaine, benzocaine, lidocaine or bupivacaine at various concentration). Isometric contracture tension was recorded throughout the experiment. The ratios of the maximal contracture tension (C-ratio) and the elapsed time (T-ratio) of the muscles treated with LA compared to those of control muscles were calculated. Tetracaine (0.125-1.0 mM) specifically reduced the C-ratio. Procaine (0.5-1.0 mM) and tetracaine (10-60 microM) increased the T-ratio. Procaine (8-16 mM), benzocaine (4-8 mM), lidocaine (0.5-4 mM) and bupivacaine (0.125-1 mM) reduced the T-ratio. The influences of LAs on ryanodine-induced contracture could be explained in terms of their effects on the Ca(2+)-induced Ca2+ release mechanism, direct Ca2+ efflux from sarcoplasmic reticulum (SR), activity of Ca2+ uptake into SR and ryanodine-receptor binding. The complexity of LA effects on ryanodine-induced contracture will affect the results of ryanodine contracture tests for malignant hyperthermia when the muscle specimen is excised under local anesthesia.

Transient expression of [D-Ala2] deltorphin I-like immunoreactivity in prenatal rat small intestine.

We studied the distribution of immunoreactive elements for [D-Ala2] deltorphin I (DADTI), a delta-opioid receptor ligand, in fetal and postnatal rat small intestine. DADTI-like immunoreactive cells were detected transiently on embryonic Days 20 and 21. Electron microscopic examination revealed that positive staining occurred in mucous epithelial cells, either mature goblet cells or undifferentiated cells containing only a few mucous granules. Positive immunoreaction products in mature goblet cells were confined in their apical cytoplasm to the luminal parts of mucous granule aggregates. The result suggests that a DADTI-like molecule(s) is synthesized in rat intestinal goblet cells and is secreted in a diacrine fashion into the intestinal lumen at a late fetal period. The molecule(s) thus secreted may be important for the intestine of rats just before birth, because DADTI-like immunopositive goblet cells are no longer seen at any postnatal period.